JP6857293B2 - Cryopreservation container - Google Patents

Description

The present invention relates to a cryopreservation container, and more particularly to a cryopreservation container for cryopreserving biological samples such as animal sperms, fertilized eggs, and germ cells.

As a cryopreservation container for cryopreserving biological samples such as animal sperms, fertilized eggs, and germ cells, an inner tank having an upper entrance for taking in and out a frozen object and an inner tank surrounding the inner tank. A triple structure including an intermediate tank that is arranged and forms a liquid nitrogen storage portion between the inner tank and an outer tank that is arranged so as to surround the intermediate tank and forms a heat insulating layer between the intermediate tank and the intermediate tank. Containers are known (see, for example, Patent Document 1).

Japanese Unexamined Patent Publication No. 2001-141346

The triple-structured cryopreservation container described in Patent Document 1 has an advantage that a frozen object such as a biological sample is not contaminated by liquid nitrogen, but is stored in a liquid nitrogen storage section. Since the peripheral wall of the inner tank is cooled to about −190 ° C. by liquid nitrogen, the air in the inner tank may condense on the inner surface of the peripheral wall to become liquid air, which may accumulate at the bottom of the inner tank in the process of long-term operation. If the amount of this liquid air increases, it may come into contact with the object to be frozen. Therefore, it is necessary to periodically remove the liquid air from the inner tank. In order to remove the liquid air from the inner tank, it is necessary to take out the object to be frozen stored in the inner tank, which not only has a problem that the temperature of the object to be frozen rises, but also requires a great deal of time and effort. Was.

Therefore, an object of the present invention is to provide a cryopreservation container having a structure in which liquid air does not accumulate in the inner tank.

In order to achieve the above object, the cryopreservation container of the present invention is arranged so as to surround the inner tank that forms a storage portion for the object to be frozen, and forms a liquid nitrogen storage portion between the inner tank. A triple-structured freezing that is provided with an intermediate tank to be used and an outer tank that is arranged so as to surround the intermediate tank and forms a heat insulating layer between the intermediate tank and an entrance / exit for taking in and out an object to be frozen. In the storage container, a drainage member having an upper end that penetrates the inner tank and opens to the bottom of the inner tank and a lower end that protrudes into the heat insulating layer is provided, and a drainage member is provided at the lower end of the heat insulating layer. It is characterized in that one end of the evaporation / discharge pipe arranged in the above is connected and the other end of the evaporation / discharge pipe is opened to the outside of the outer tank.

Further, in the cryopreservation container of the present invention, the evaporation / discharge pipe has an ascending slope from one end connected to the drainage member toward the other end open to the outside, or is opposite to the connection portion to the drainage member. It is characterized by having a stop valve.

Further, the evaporation / discharge pipe is arranged in a spiral spiral, meandering or polygonal multi-layer winding shape in a plan view in the heat insulating layer below the intermediate tank from one end connected to the drainage member. It is characterized in that it rises upward on the outer peripheral side from below the intermediate tank, penetrates the outer tank, and protrudes to the outside.

According to the cryopreservation container of the present invention, the liquid air generated in the inner tank can be automatically discharged to the outside, so that the object to be frozen is not contaminated and the liquid air can be removed from the inner tank. No work is required, and the object to be frozen can be kept in a predetermined frozen state for a long period of time.

It is a partial cross-sectional perspective view which shows one form example of the cryopreservation container of this invention. It is also a front view. It is also a plan view. FIG. 3 is a sectional view taken along line IV-IV of FIG. FIG. 3 is a sectional view taken along line VV of FIG. FIG. 2 is a sectional view taken along line VI-VI of FIG.

1 to 6 show an example of a form of the cryopreservation container of the present invention. The cryopreservation container shown in this embodiment has a triple structure having an inner tank 11, an intermediate tank 12 arranged so as to surround the inner tank 11, and an outer tank 13 arranged so as to surround the intermediate tank 12. Is formed of. The inner tank 11 is a storage unit 14 for storing the object to be frozen in a predetermined frozen state, and a liquid nitrogen storage for storing liquid nitrogen between the inner tank 11 and the intermediate tank 12 is provided. The portion 15 is formed. Further, a heat insulating layer 16 adopting the laminated vacuum method is formed between the intermediate tank 12 and the outer tank 13.

In the upper center of the inner tank 11, the intermediate tank 12, and the outer tank 13, entrances 11a, 12a, 13a for taking in and out the object to be frozen are provided in the storage portion 14, and the entrance 12a and the outside of the intermediate tank 12 are provided. A connecting cylinder 17 for airtightly connecting both the entrances 12a and 13a is provided between the entrance and exit 13a of the tank 13, and a heat insulating lid 18 having heat insulating properties is detachably inserted inside the connecting cylinder 17. Has been done. Further, the bottom plate of the intermediate tank 12 is provided with a circular opening 12b, and the central portion 11b of the bottom plate of the inner tank 11 is fitted into the opening 12b and airtightly joined to the inner tank 11. The bottom plate central portion 11b is in contact with the heat insulating layer 16.

The bottom plate central portion 11b of the inner tank 11 penetrates the bottom plate central portion 11b, the upper end opens to the lowermost lower end of the bottom surface of the inner tank 11, and the lower end is a bottomed tubular drainage liquid arranged in the heat insulating layer 16. A member 19 is provided, and the lower end of the drainage member 19 is held by a heat insulating support member 20 that supports the inner tank 11 and the intermediate tank 12 via the drainage member 19 at the center portion 13b of the bottom plate of the outer tank 13. There is.

Further, one end of the evaporation / discharge pipe 21 communicating with the inside of the cylinder of the drainage member 19 is connected to the side surface of the lower end portion of the drainage member 19. The evaporation / discharge pipe 21 is arranged in the heat insulating layer 16, and is a spiral portion 21a formed spirally in the lower portion of the intermediate tank 12 centering on one end connected to the drainage member 19 in a plan view. And a rising portion 21b that rises vertically upward from the end of the spiral portion 21a located on the outer peripheral side of the lower portion of the intermediate tank 12, and the upper end portion of the rising portion 21b is outside via the penetrating member 22. It protrudes to the outside by penetrating the top plate 13c of the tank 13. The base end of the inverted J-shaped tube 23 having an inverted J-shape and the tip opening downward at the lower side of the container is connected to the other end of the evaporation / discharge tube 21 protruding to the outside. Further, the spiral portion 21a of the evaporation / discharge pipe 21 has an ascending slope toward the end of the spiral portion 21a with one end connected to the drainage member 19 as the lowermost end.

On the top plate 14c of the outer tank 13, in addition to the inverted J-shaped tube 23 provided at the other end of the evaporation / discharge tube 21, a pair of left and right handles 24 and 24 and liquid nitrogen in the liquid nitrogen storage section 15 are charged. A liquid nitrogen filling pipe 25 for filling, a liquid level sensor 26 for monitoring the liquid level of liquid nitrogen in the liquid nitrogen storage section 15, a vacuum exhaust port 27 for vacuum exhausting the inside of the heat insulating layer 16 and the like. An outer lid 28 that covers the upper part of the heat insulating lid 18 so as to be openable and closable is provided. The filling pipe 25a of the liquid nitrogen filling pipe 25 and the protective pipe 26a of the liquid level sensor 26 penetrate the outer peripheral portion of the top plate of the outer tank 13 and the outer peripheral portion of the top plate of the intermediate tank 12 via the penetrating members 25b and 26b to liquid. It is inserted into each of the nitrogen storage portions 15.

In this cryopreservation container, an inner tank 11 to which a drainage member 19 is attached, an intermediate tank 12, an outer tank 13 and the like are combined in a predetermined state, and the inside of the heat insulating layer 16 is evacuated from the vacuum exhaust port 27 by a vacuum pump to insulate. After the state is set, a predetermined amount of liquid nitrogen is filled into the liquid nitrogen storage unit 15 from the liquid nitrogen filling pipe 25 and used.

After the atmosphere of the storage unit 14 in the inner tank 11 cooled by the liquid nitrogen in the liquid nitrogen storage unit 15 becomes a predetermined low temperature state, the outer lid 28 is opened, the heat insulating lid 18 is pulled out, and the inside of the storage unit 14 Store the object to be frozen in. The frozen object can be taken in and out of the frozen object storage portion 14 in the same manner as in the conventional case depending on the shape of the frozen object and the like. During the cryopreservation of the object to be frozen, liquid nitrogen is appropriately replenished from the liquid nitrogen filling pipe 25 according to the liquid level of the liquid nitrogen in the liquid nitrogen storage unit 15 detected by the liquid level sensor 26.

The air in the storage unit 14 is cooled to about −190 ° C. by the liquid nitrogen in the liquid nitrogen storage unit 15 during use in which the object to be frozen is stored in the storage unit 14 and stored frozen. Condensation on the wall surface of the tank 11 becomes liquid air, and when it flows down to the bottom surface of the inner tank 11 along the wall surface of the inner tank 11, it flows into the cylinder of the drainage member 19 opened at the lowermost end of the bottom surface, and further evaporates. It flows into the inside of the discharge tube 21.

Since the evaporation / discharge pipe 21 is arranged in the heat insulating layer 16, the temperature is higher than the boiling point (about −190 ° C.) of the liquid air, so that the liquid air flowing into the evaporation / discharge pipe 21 is It gradually vaporizes into gaseous air, rises along the upslope of the spiral portion 21a, passes through the rising portion 21b, passes through the inverted J-shaped tube 23 connected to the protruding portion 21c, and is discharged downward to the outside air. To. At this time, since the spiral portion 21a of the evaporation / discharge pipe 21 has an upward gradient, the gaseous air vaporized in the evaporation / discharge pipe 21 does not flow back toward the drainage member 19, and the drainage member does not flow back. The liquid air flowing down into the 19 is not blown up by the gaseous air.

Therefore, since the liquid air generated in the inner tank 11 can be discharged from the drain member 19 to the outside through the evaporation / discharge pipe 21, the liquid air is not accumulated in the inner tank 11 and the liquid air is covered. It is possible to avoid contact with frozen matter. This makes it possible to prevent the object to be frozen from being contaminated by liquid air. Further, since the work of removing the liquid air from the inner tank 11 becomes unnecessary, the object to be frozen can be stored and stored in the low temperature storage portion 14 in the inner tank 11 in a stable frozen state for a long period of time. ..

Each tank and pipe can be formed of a metal such as an aluminum alloy or stainless steel, and the structure and shape can be appropriately set according to conditions such as a storage capacity. Further, the filling pipe 25a and the protective pipe 26a can be combined into one penetrating member so that the shoulder portion of the outer tank 13 and the shoulder portion of the intermediate tank 12 can be penetrated at one place, and an appropriate heat insulating structure shall be adopted. You can also.

Further, the evaporation / discharge pipe 21 preferably has a structure capable of accumulating an appropriate amount of liquid air in the pipe, and as shown in the above embodiment, the evaporative discharge pipe 21 rises from the connection portion to the drainage member 19 toward the rising portion 21b. By providing the spiral portion 21a with a gradient, liquid air can be stored in the vicinity of the drainage member 19 on the lower side of the spiral portion 21a. The shape of the pipe from the connection portion to the drainage member 19 to the rising portion 21b is not limited to the spiral shape, but the liquid air can be stored by meandering, polygonal multi-layer winding shape, circular shape, or the like. Can be formed to do so. Further, it can be formed so as to store liquid air by providing a tank-shaped large-diameter pipe in the middle of the pipe from the connection portion to the drainage member 19 to the rising portion 21b.

Further, the rising portion 21b may not be provided up to the upper part of the outer tank 13, but may be projected to the outside of the outer tank 13 at the vertical intermediate portion of the outer tank 13, and the protruding portion may be formed instead of the inverted U-shaped pipe 23. A check valve may be provided on the 21c. The position of the drainage member 19 for discharging the liquid air from the inner tank 11 is also arbitrary, and it can be provided at an arbitrary position where the liquid air can be discharged from the inner tank 11, and the shape and the number of installations are also arbitrary. For example, liquid air can be discharged even if the end portion of the rising portion 21b is opened on the wall surface rising from the bottom surface of the outer tank 13. Further, the drainage member 19 and the evaporation / discharge pipe 21 can be integrally formed. Further, the liquid nitrogen storage unit 15 may be filled with a liquid nitrogen absorber. As a result, when the cryopreservation container is moved, it is possible to prevent a trouble that liquid nitrogen wraps around the storage portion 14 along the peripheral wall of the intermediate tank 12. Further, all the pipes from the connection portion of the evaporation / discharge pipe 21 to the drainage member 19 to the rising portion 21b do not have to have an uphill slope. For example, it may be horizontal, or even if only a part of the slope is uphill, it is possible to prevent the gaseous air vaporized in the evaporation / discharge pipe 21 from flowing back toward the drainage member 19. Further, a check valve can also be provided at the connection portion of the evaporation / discharge pipe 21 to the drainage member 19, to obtain a check flow prevention effect.

The shape and size of the object to be frozen to be cryopreserved in the cryopreservation container are arbitrary. For example, a biological sample enclosed in a vial or an ampoule or various samples molded into a sheet can be targeted. It suffices to ensure that the object to be frozen is taken in and out of the container and the object to be frozen is held in the storage unit 14, and usually, it may be performed in the same manner as in the conventional case.

11 ... Inner tank, 11a ... Doorway, 11b ... Bottom plate center, 12 ... Intermediate tank, 12a ... Doorway, 12b ... Opening, 13 ... Outer tank, 13a ... Doorway, 13b ... Bottom plate center, 14 ... Storage part, 15 ... Liquid nitrogen storage, 16 ... Insulation layer, 17 ... Connecting cylinder, 18 ... Insulation lid, 19 ... Drainage member, 20 ... Insulation support member, 21 ... Evaporation / discharge pipe, 21a ... Spiral part, 21b ... Rise Part, 21c ... Protruding part, 22 ... Penetrating member, 23 ... Discharge tube, 24 ... Handle, 25 ... Liquid nitrogen filling tube, 25a ... Filling tube, 25b ... Penetrating member, 26 ... Liquid level sensor, 26a ... Protective tube, 26b ... Penetration member, 27 ... Vacuum exhaust port, 28 ... Outer lid

Claims (3)

An inner tank that forms a storage portion for the object to be frozen and an intermediate tank that is arranged so as to surround the inner tank and forms a liquid nitrogen storage portion between the inner tank and the intermediate tank are arranged so as to surround the intermediate tank. In a triple-structured cryopreservation container provided with an outer tank that forms a heat insulating layer between the intermediate tank and an upper portion for taking in and out an object to be frozen, the upper end penetrates the inner tank and the upper end is inside. A drainage member that opens at the bottom of the tank and whose lower end protrudes into the heat insulating layer is provided, and one end of an evaporation / discharge pipe arranged in the heat insulating layer is connected to the lower end of the drainage member. A cryopreservation container characterized in that the other end of the evaporation / discharge tube is opened to the outside of the outer tank. The evaporation / discharge pipe is characterized by having an ascending slope from one end connected to the drainage member toward the other end opening to the outside, or a check valve at a connection portion to the drainage member. The cryopreservation container according to claim 1. The evaporation / discharge pipe is arranged in a spiral, meandering, or polygonal multi-layer winding shape in a plan view in the heat insulating layer below the intermediate tank from one end connected to the drainage member. The cryopreservation container according to claim 1 or 2, wherein the container rises upward on the outer peripheral side from below, penetrates the outer tank, and protrudes to the outside.

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